Michael Lein

Publications (101)390.11 Total impact

• Article: N'-Formyl-2-(5-nitrothiophen-2-yl)benzothiazole-6-carbohydrazide as a potential anti-tumour agent for prostate cancer in experimental studies.

  Juan R Rodrigues, Jaime Charris, José Camacho, Arthur Barazarte, Neira Gamboa, Bianca Nitzsche, Michael Höpfner, Michael Lein, Klaus Jung, Claudia Abramjuk
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  ABSTRACT: Benzothiazoles (BZTs) represent organic compounds with different biological actions. In this study we aimed to investigate ten newly synthesized BZT derivatives as potential anti-tumour agents against prostate cancer in vitro and in vivo. The cytotoxic effect of these compounds was screened on the human prostate cancer cell lines PC-3 and LNCaP. The most effective compound, N'-formyl-2-(5-nitrothiophen-2-yl)benzothiazole-6-carbohydrazide, was further characterized regarding its dose- and time-dependent effects on cell viability and proliferation (XTT test) as well as on adhesion and spreading (real-time cell analyzer xCelligence), migration (scratch-wound repair assay) and invasion (Boyden chamber) of the cells. This BZT derivative was also tested as an inhibitor of angiogenesis (chicken chorioallantoic membrane assay), clonogenic activity (soft agar) and matrix metalloproteinase 9 (gelatin zymography). N'-Formyl-2-(5-nitrothiophen-2-yl)benzothiazole-6-carbohydrazide significantly inhibited all tested properties of the prostate cancer cell lines and showed low toxic in vitro and in vivo effects. The in vitro anti-tumour activity of this compound was confirmed by the in vivo effects on PC-3 xenografts in nude mice. Tumour growth was decreased in treated compared with untreated mice. These results suggest the potential capacity of BZTs and in particular N'-formyl-2-(5-nitrothiophen-2-yl)benzothiazole-6-carbohydrazide as anti-tumour agents for the treatment of prostate cancer.
  The Journal of pharmacy and pharmacology. 03/2013; 65(3):411-22.
• Article: Value of Prostate Specific Antigen Density and Percent Free Prostate Specific Antigen for Prostate Cancer Prognosis.

  Jonas Busch, Kristin Hamborg, Hellmuth-Alexander Meyer, John Buckendahl, Ahmed Magheli, Michael Lein, Klaus Jung, Kurt Miller, Carsten Stephan
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  ABSTRACT: PURPOSE: Limited data exist on the relationship of percent free prostate specific antigen and prostate specific antigen density with prostate cancer prognosis. Therefore, we compared percent free prostate specific antigen and prostate specific antigen density with prostate specific antigen, Gleason sum and stage to predict prostate cancer prognosis in a large cohort using a single prostate specific antigen and free prostate specific antigen assay. MATERIALS AND METHODS: Between 1999 and 2007 a total of 1,656 patients with prostate cancer underwent laparoscopic radical prostatectomy at the Charité Berlin. There were 322 patients excluded from analysis for a variety of reasons. The final 1,334 patients had prostate specific antigen, free prostate specific antigen, prostate volume and complete pathological analysis available. RESULTS: Median followup was 60.3 months (range 0.2 to 135). Median age (63 years, range 43 to 75) did not differ between the 1,092 patients without and the 242 with biochemical recurrence (p = 0.956), but prostate volume, prostate specific antigen and percent free prostate specific antigen differed significantly (p <0.0001). While prostate specific antigen and prostate specific antigen density increased significantly in patients with Gleason less than 7, 7 and greater than 7 tumors, percent free prostate specific antigen decreased significantly (p <0.0001). Prostate specific antigen, percent free prostate specific antigen and prostate specific antigen density differed significantly between pT2 and pT3 tumors, and between patients with vs without positive surgical margins. On univariate analysis Gleason sum, pathological stage, positive surgical margin, total prostate specific antigen, percent free prostate specific antigen and prostate specific antigen density were predictors of biochemical recurrence-free survival. Multivariate Cox regression analysis identified Gleason sum, pathological stage, positive surgical margin and prostate specific antigen density as independent predictors of biochemical recurrence-free survival, while percent free prostate specific antigen and total prostate specific antigen failed to be significant. CONCLUSIONS: Few models for prostate cancer prognosis include prostate specific antigen density. There is substantial value in prostate specific antigen density but not in percent free prostate specific antigen for improving prostate cancer prognosis and biochemical recurrence prediction.
  The Journal of urology 10/2012; · 4.02 Impact Factor
• Article: Long-term oncological and continence outcomes after laparoscopic radical prostatectomy: a single-centre experience.

  Jonas Busch, Carsten Stephan, Anja Herold, Barbara Erber, Carsten Kempkensteffen, Stefan Hinz, Michael Lein, Steffen Weikert, Kurt Miller, Ahmed Magheli
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  ABSTRACT: Study Type - Therapy (case series) Level of Evidence 4 What's known on the subject? and What does the study add? Over the past decade, minimally invasive laparoscopic radical prostatectomy and more recently robot-assisted laparoscopic prostatectomy have been introduced and have proven equally effective compared with open surgery in terms of mid-term cancer control and complication rates. Because long-term data is lacking, open prostatectomy is still considered the 'gold standard' by some authors, who argue that minimally invasive approaches have to measure up to the excellent long-term results of open surgery. This study represents one of the largest series (1845 patients) of minimally invasive radical prostatectomy with extended follow-up (11.3 years) and detailed data on oncological outcome and postoperative incontinence. It therefore supplies previously lacking information on these details for minimally invasive prostate surgery and provides important information for patient counselling. OBJECTIVE: •  To investigate biochemical recurrence (BCR) rates and data on postoperative incontinence in a large laparoscopic radical prostatectomy (LRP) cohort with extended follow-up. MATERIALS AND METHODS: •  BCR and independent predictors of BCR were identified using Kaplan-Meier and Cox regression analysis of 1845 patients who underwent LRP from 1999 to 2007. •  Urinary incontinence was evaluated by pads per day and stratified as follows: 0-1 pad: no incontinence; 2-3 pads: mild incontinence; and ≥3 pads: severe incontinence. RESULTS: •  Organ-confined disease, extraprostatic extension, seminal vesicle invasion and lymph node metastasis were present in 71.3%, 20.5%, 6.7% and 3.2% of patients, respectively. The positive surgical margin rate was 29.2%. •  Postoperatively, 74.9% of the patients were continent, while 9.2% had mild and 15.9% severe incontinence. •  The mean follow-up was 5 years with a maximum follow-up of 11.3 years. •  There were 51 overall deaths and six deaths from prostate cancer. The 5-year, 8-year and 10-year BCR-free survival rates were 83.9%, 78.6% and 75.6%, respectively. •  On univariate analyses preoperative D'Amico risk classification, pathological tumour stage, postoperative Gleason sum and surgical margin status were predictors of BCR (P < 0.001). •  On multivariable analysis, D'Amico classification, Gleason sum (P < 0.001), postoperative tumour stage (P < 0.001), nodal status (P < 0.001) and surgical margin status (P= 0.002) were independent predictors of BCR. CONCLUSIONS: •  LRP offers excellent long-term functional and oncological results with a low incidence of BCR for patients with localized disease. •  These results could be used for patient counselling before robot-assisted laparascopic prostatectomy (RALP) until long-term follow-up data for RALP is available.
  BJU International 06/2012; · 2.84 Impact Factor
• Article: Impact of positive surgical margins on oncological outcome following laparoscopic radical prostatectomy (LRP): long-term results.

  Jonas Busch, Carsten Stephan, Annett Klutzny, Stefan Hinz, Carsten Kempkensteffen, Ergin Kilic, Michael Lein, Steffen Weikert, Kurt Miller, Ahmed Magheli
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  ABSTRACT: PURPOSE: The impact of positive surgical margins (PSM) on biochemical recurrence (BCR) has been heavily debated in laparoscopic radical prostatectomy (LRP). The aim of this study was to investigate the impact of PSM on BCR following LRP in patients with extended follow-up. METHODS: Retrospective chart review of 1,845 patients who underwent LRP from 1999 to 2007. Predictors of PSM and BCR were identified utilizing univariate and multivariable logistic and Cox regression analyses, respectively. RESULTS: Five hundred and thirty-seven patients (29.1 %) had a PSM. Median postoperative follow-up was 56 months. 10-year BCR-free survival was 59.2 and 82.9 % for patients with and without PSM, respectively (p < 0.0001). Clinical stage T2 (OR 1.66; CI 1.23-2.25; p = 0.001), a biopsy Gleason sum > 7 (OR 1.84; CI 1.06-3.18; p = 0.031) and preoperative prostate-specific antigen (PSA) levels of 10-20 ng/mL (OR 1.58; CI 1.12-2.23; p = 0.010) and >20 ng/mL (OR 6.82; CI 3.51-13.27; p < 0.0001) were independent predictors of PSM. Prostate size was inversely associated with PSM (OR 0.99; CI 0.98-1.00; p = 0.002). On multivariable analysis, LRP Gleason score of 7 (HR 2.45; CI 1.67-3.40; p < 0.0001) and >7 (HR 4.76; CI 3.15-7.19; p < 0.0001), PSM (HR 1.49; CI 1.14-2.00; p = 0.003), advanced pathological stages (p < 0.001), and PSA 10-20 ng/mL (HR 1.46; CI 1.13-1.89; p = 0.004) were independent predictors of BCR. CONCLUSIONS: We demonstrated the independent predictive value of PSM for BCR in our LRP cohort with extended follow-up. Our results could potentially be transferred to robotic RP, in which long-term follow-up is lacking.
  World Journal of Urology 05/2012; · 2.41 Impact Factor
• Article: Assay-dependent abnormalities in measurements of prostate-specific antigen in serum: an occasional occurrence, but of clinical significance.

  Klaus Jung, Carsten Stephan, Michael Lein
  Clinical Chemistry and Laboratory Medicine 03/2012; 50(3):585-6. · 2.15 Impact Factor
• Article: Effect of quinolinyl acrylate derivatives on prostate cancer in vitro and in vivo.

  Juan R Rodrigues, Jaime Charris, Rosa Ferrer, Neira Gamboa, Jorge Angel, Bianca Nitzsche, Michael Hoepfner, Michael Lein, Klaus Jung, Claudia Abramjuk
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  ABSTRACT: Quinolines and acrylates are chemical compounds which were previously described as potential antitumor agents. In this study, a series of seven new quinolinyl acrylate derivatives were synthesized and evaluated against human prostate cancer cells PC-3 and LNCaP in vitro and in vivo. The most effective compound (E)-methyl 2-(7-chloroquinolin-4-ylthio)-3-(4 hydroxyphenyl) acrylate reduced the viability in both cell lines in a time- and dose-dependent manner. Inhibitory effects were also observed on the adhesion, migration, and invasion of the prostate cancer cells as well as on the neoangiogenesis, clonogenic and MMP-9 activity. The effect in vivo was studied in PC-3 xenografts in nude mice. The results were concordant with the in vitro effects and showed decreased tumor growth in treated animals compared to controls. The study suggests the multi-target efficacy of the quinolinyl derivate against human prostate cancer cells and supports its potential therapeutic usefulness.
  Investigational New Drugs 07/2011; 30(4):1426-33. · 3.36 Impact Factor
• Article: Impact of fast-track postoperative care on intestinal function, pain, and length of hospital stay after laparoscopic radical prostatectomy.

  Ahmed Magheli, Nina Knoll, Michael Lein, Stefan Hinz, Carsten Kempkensteffen, Oliver Gralla
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  ABSTRACT: Postoperative recovery of intestinal function, ability to ambulate, and effective pain management are main features to establish an effective fast-track surgery model. We investigated pain scores, ambulation rate, and recovery of intestinal function in a cohort of patients who were undergoing laparoscopic radical prostatectomy (LRP). Fifty patients who underwent LRP in our institution were randomized to receive either conventional or fast-track postoperative care. Postoperative intestinal function was quantified by clinical signs of intestinal motility. Ambulation data were collected by means of step-count devices. Pain scores were measured by a visual analog scale. Overall satisfaction and additional measures to describe patient satisfaction with the clinical course were used as quality-of-life variables. Fast-track patients had significantly earlier propulsive intestinal motility without increased intestinal complications. Enforced mobilization led to a significantly shorter period to first deflation/defecation. Despite significantly increased ambulation rates in the fast-track group, these patients reported significantly less pain sentience during a significantly shorter hospital stay. Overall satisfaction was significantly higher in the fast-track cohort during the hospital stay. With the implementation of fast-track concepts for LRP, patients can be discharged to home earlier with fewer complications, lower pain scores, and an overall higher satisfaction with life.
  Journal of endourology / Endourological Society 07/2011; 25(7):1143-7. · 1.75 Impact Factor
• Article: Between-method differences in prostate-specific antigen assays affect prostate cancer risk prediction by nomograms.

  Carsten Stephan, Kerstin Siemssen, Henning Cammann, Frank Friedersdorff, Serdar Deger, Mark Schrader, Kurt Miller, Michael Lein, Klaus Jung, Hellmuth-Alexander Meyer
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  ABSTRACT: To date, no published nomogram for prostate cancer (PCa) risk prediction has considered the between-method differences associated with estimating concentrations of prostate-specific antigen (PSA). Total PSA (tPSA) and free PSA were measured in 780 biopsy-referred men with 5 different assays. These data, together with other clinical parameters, were applied to 5 published nomograms that are used for PCa detection. Discrimination and calibration criteria were used to characterize the accuracy of the nomogram models under these conditions. PCa was found in 455 men (58.3%), and 325 men had no evidence of malignancy. Median tPSA concentrations ranged from 5.5 μg/L to 7.04 μg/L, whereas the median percentage of free PSA ranged from 10.6% to 16.4%. Both the calibration and discrimination of the nomograms varied significantly across different types of PSA assays. Median PCa probabilities, which indicate PCa risk, ranged from 0.59 to 0.76 when different PSA assays were used within the same nomogram. On the other hand, various nomograms produced different PCa probabilities when the same PSA assay was used. Although the ROC curves had comparable areas under the ROC curve, considerable differences were observed among the 5 assays when the sensitivities and specificities at various PCa probability cutoffs were analyzed. The accuracy of the PCa probabilities predicted according to different nomograms is limited by the lack of agreement between the different PSA assays. This difference between methods may lead to unacceptable variation in PCa risk prediction. A more cautious application of nomograms is recommended.
  Clinical Chemistry 05/2011; 57(7):995-1004. · 7.91 Impact Factor
• Article: Sarcosine in prostate cancer tissue is not a differential metabolite for prostate cancer aggressiveness and biochemical progression.

  Florian Jentzmik, Carsten Stephan, Michael Lein, Kurt Miller, Beate Kamlage, Bianca Bethan, Glen Kristiansen, Klaus Jung
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  ABSTRACT: Sarcosine in prostate cancer tissue samples was recently reported to be increased during prostate cancer progression to metastasis and suggested to be a key metabolite of cancer cell invasion and aggressiveness. We reevaluated sarcosine in prostate cancer tissue samples as a potential indicator of tumor aggressiveness, and as a predictor of recurrence-free survival. Sarcosine in matched samples of malignant and nonmalignant tissue from 92 patients with prostate cancer after radical prostatectomy was measured in the framework of a global metabolite profiling study of prostate cancer by gas chromatography/mass spectrometry. We related results to age, prostate volume, tumor stage, Gleason score, preoperative prostate specific antigen and biochemical recurrence, defined as a persistent prostate specific antigen increase of greater than 0.2 ng/ml. Nonparametric statistical tests, ROC curves and Kaplan-Meier analyses were done. Median sarcosine content in tissue was about 7% higher in matched malignant vs nonmalignant samples, which was significantly. Sarcosine values were not associated with tumor stage (pT2 vs pT3), tumor grade (Gleason score less than 7 vs 7 or greater) or biochemical recurrence. The lack of metastatic tissue samples was a study limitation. Sarcosine in prostate cancer tissue samples cannot be considered a suitable predictor of tumor aggressiveness or biochemical recurrence.
  The Journal of urology 02/2011; 185(2):706-11. · 4.02 Impact Factor
• Article: Bone turnover markers as predictors of mortality risk in prostate cancer patients with bone metastases following treatment with zoledronic acid.

  Klaus Jung, Kurt Miller, Manfred Wirth, Michael Albrecht, Michael Lein
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  ABSTRACT: Clinical data have limited validity for predicting the survival of prostate cancer (PCa) patients with bone metastases. There is a need to improve the predictive evidence both for clinicians and patients. To evaluate the predictive ability of serum bone markers for mortality risk in PCa patients with bone metastases. We conducted a survival analysis in relation to bone markers in a subgroup of 52 patients treated with zoledronic acid (4 mg every 4 wk for 15 mo) in a prospective, multicentre trial during 2002-2005, about 4 yr after the end of the trial. Serum levels of total and bone-specific alkaline phosphatase, amino-terminal procollagen propeptides of type I collagen (PINP), cross-linked N-terminal (NTx) and cross-linked C-terminal telopeptides of type I collagen (ICTP), C-terminal telopeptides of type I collagen, prostate-specific antigen from the last visit of the treatment study, and clinical data were related to the overall survival (OS) status of patients in the follow-up. Univariate and multivariate Cox regression analyses with internal bootstrapping validation and concordance index calculations were performed. Out of the 52 patients followed, 34 died within a median follow-up of 13.8 mo, and 18 patients were alive at a median follow-up of 43.8 mo. The patients who died within the follow-up period had significantly higher concentrations of ICTP, NTx, and PINP than the surviving patients. Cox regression models with clinical data and bone markers showed that ICTP and PINP were most predictive for mortality risk in addition to the occurrence of skeletal-related complications and the continuation of treatment with zoledronic acid. Internal validation confirmed the reliability of the results, although the sample size was small. PINP and ICTP can be considered suitable predictors for the OS of PCa patients with bone metastases.
  European urology 12/2010; 59(4):604-12. · 7.67 Impact Factor
• Article: New 4-maleamic acid and 4-maleamide peptidyl chalcones as potential multitarget drugs for human prostate cancer.

  Juan Rodrigues, Claudia Abramjuk, Luis Vásquez, Neira Gamboa, José Domínguez, Bianca Nitzsche, Michael Höpfner, Radostina Georgieva, Hans Bäumler, Carsten Stephan, Klaus Jung, Michael Lein, Anja Rabien
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  ABSTRACT: The objective of this study was to investigate the effect of new 4-maleamic acid and 4-maleamide peptidyl chalcone derivatives against human prostate cancer in vitro and in vivo. From a series of 21 chalcones, the effects of the three best inhibitors of PC-3 and LNCaP cell viability on growth, including cell cycle changes, adhesion, migration, and cell invasion, as well as their ability to inhibit angiogenesis, clonogenic activity, and matrix metalloproteinases MMP-2 and MMP-9, were tested. The effects in vivo were studied in PC-3 and LNCaP xenografts. Three of the examined chalcones reduced cell viability in both cell lines in a strong dose- and time-dependent manner. An inhibition of the cell cycle progress was observed. These changes were accompanied with the inhibition of cell adhesion, migration, and invasion as well as with reduced neovascularization in chick embryos, tumor colony formation, and MMP-9 activity. The in vivo results demonstrated the strong activity of these structures as inhibitors of tumor development in nude mice compared to non-treated animals. The results suggest the multitarget efficacy of 4-maleamic acid and 4-maleamide peptidyl chalcones against human prostate cancer cells and emphasize the potential therapeutic relevance of these compounds.
  Pharmaceutical Research 12/2010; 28(4):907-19. · 4.09 Impact Factor
• Article: Suitable reference genes for relative quantification of miRNA expression in prostate cancer.

  Annika Schaefer, Monika Jung, Kurt Miller, Michael Lein, Glen Kristiansen, Andreas Erbersdobler, Klaus Jung
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  ABSTRACT: Real time quantitative PCR (qPCR) is the method of choice for miRNA expression studies. For relative quantification of miRNAs, normalization to proper reference genes is mandatory. Currently, no validated reference genes for miRNA qPCR in prostate cancer are available. In this study, the expression of four putative reference genes (hsa-miR-16, hsa-miR-130b, RNU6-2, SNORD7) was examined with regard to their use as normalizer. After SNORD7 was already shown an inappropriate reference gene in preliminary experiments using total RNA pools, we studied the expression of the putative reference genes in tissue and normal adjacent tissue sample pairs from 76 men with untreated prostate carcinoma collected after radical prostatectomy. hsa-miR-130b and RNU6-2 showed no significantly different expression between the matched malignant and non-malignant tissue samples, whereas hsa-miR-16 was significantly underexpressed in malignant tissue. Softwares geNorm and Normfinder predicted hsa- miR-130b and the geometric mean of hsa-miR-130b and RNU6-2 as the most stable reference genes. Normalization of the four miRNAs hsa-miR-96, hsa- miR-125b, hsa-miR-205, and hsa-miR-375, which were previously shown to be regulated, shows that normalization to hsa-mir-16 can lead to biased results. We recommend using hsa-miR-130b or the geometric mean of hsa-miR-130b and small RNA RNU6-2 for normalization in miRNA expression studies of prostate cancer.
  Experimental and Molecular Medicine 10/2010; 42(11):749-58. · 2.48 Impact Factor
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  Article: Sarcosine in urine after digital rectal examination fails as a marker in prostate cancer detection and identification of aggressive tumours.

  Florian Jentzmik, Carsten Stephan, Kurt Miller, Mark Schrader, Andreas Erbersdobler, Glen Kristiansen, Michael Lein, Klaus Jung
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  ABSTRACT: Sarcosine in urine was recently suggested to be a promising tool in prostate cancer (PCa) diagnostics. To reevaluate sarcosine as a potential biomarker for early PCa detection and for prediction of tumour aggressiveness. Sarcosine was measured in urine samples from 106 PCa patients and 33 patients with no evidence of malignancy (NEM), confirmed by 8-12 core prostate biopsies, after standardised digital rectal examination, as well as from 12 healthy men and women. The results were related to the clinicopathologic data on prostate volume, tumour stage, Gleason score, and prostate specific antigen (PSA). Sarcosine in urine was determined by gas chromatography-mass spectrometry using a commercial amino acid assay and was normalised to urine creatinine. Nonparametric statistical tests and receiver operating characteristics (ROC) analyses were performed to assess the diagnostic performance. The median sarcosine-creatinine ratio in urine was 13% lower in PCa than in NEM patients. Sarcosine values were not associated with tumour stage (pT2 vs pT3) or grade (Gleason score <7 vs > or = 7). ROC analyses proved that the discrimination between PCa and NEM patients was not improved by sarcosine in comparison with total PSA, but it was significantly worse than the percent free PSA. The higher proportion of PCa than NEM patients can be considered a limitation of this study. Sarcosine in urine after rectal digital examination cannot be considered as a suitable marker to differentiate between patients with and without PCa.
  European urology 07/2010; 58(1):12-8; discussion 20-1. · 7.67 Impact Factor
• Article: Diagnostic and prognostic value of T-cell receptor gamma alternative reading frame protein (TARP) expression in prostate cancer.

  Florian R Fritzsche, Carsten Stephan, Josefine Gerhardt, Michael Lein, Irina Hofmann, Klaus Jung, Manfred Dietel, Glen Kristiansen
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  ABSTRACT: T-cell receptor gamma chain alternative reading frame protein (TARP) has recently been proposed as being up-regulated in prostate cancer (PCA). Additionally, TARP has been proposed as a potential therapeutic target for cancer therapy. We analysed the protein expression of TARP in a large well characterised prostate cancer cohort to assess its diagnostic and prognostic value. Methodologically, we constructed a tissue microarray comprising more than 600 PCA cases including matching benign prostate tissue. TARP protein expression was carefully analysed and associated with clinico-pathological parameters, PSA-relapse free survival and expression data of established and proposed diagnostic markers (AMACR, p63, GOLPH2). Our results show that TARP is significantly over-expressed in the vast majority (approximately 85%) of PCA in comparison to non neoplastic prostate tissue. Its expression was associated with conventional markers of unfavourable and more aggressive tumour behaviour. However, a prognostic value of TARP could not be found. The diagnostic value of TARP is limited in comparison to AMACR, p63 or GOLPH2. Since TARP specific immunologic therapy regimen are currently being tested, the high frequency of TARP over-expression in PCA conveys a high potential for a predictive and potentially therapeutic use of this biomarker.
  Histology and histopathology 06/2010; 25(6):733-9. · 2.48 Impact Factor
• Article: Prevalence of TMPRSS2-ERG and SLC45A3-ERG gene fusions in a large prostatectomy cohort.

  Raquel Esgueva, Sven Perner, Christopher J LaFargue, Veit Scheble, Carsten Stephan, Michael Lein, Florian R Fritzsche, Manfred Dietel, Glen Kristiansen, Mark A Rubin
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  ABSTRACT: The majority of prostate cancers harbor recurrent gene fusions between the hormone-regulated TMPRSS2 and members of the ETS family of transcription factors, most commonly ERG. Prostate cancer with ERG rearrangements represent a distinct sub-class of tumor based on studies reporting associations with histomorphologic features, characteristic somatic copy number alterations, and gene expression signatures. This study describes the frequency of ERG rearrangement prostate cancer and three 5 prime (5') gene fusion partners (ie, TMPRSS2, SLC45A3, and NDRG1) in a large prostatectomy cohort. ERG gene rearrangements and mechanism of rearrangement, as well as rearrangements of TMPRSS2, SLC45A3, and NDRG1, were assessed using fluorescence in situ hybridization (FISH) on prostate cancer samples from 614 patients treated using radical prostatectomy. ERG rearrangement occurred in 53% of the 540 assessable cases. TMPRSS2 and SLC45A3 were the only 5' partner in 78% and 6% of these ERG rearranged cases, respectively. Interestingly, 11% of the ERG rearranged cases showed concurrent TMPRSS2 and SLC45A3 rearrangements. TMPRSS2 or SLC45A3 rearrangements could not be identified for 5% of the ERG rearranged cases. From these remaining cases we identified one case with NDRG1 rearrangement. We did not observe any associations with pathologic parameters or clinical outcome. This is the first study to describe the frequency of SLC45A3-ERG fusions in a large clinical cohort. Most studies have assumed that all ERG rearranged prostate cancers harbor TMPRSS2-ERG fusions. This is also the first study to report concurrent TMPRSS2 and SLC45A3 rearrangements in the same tumor focus, suggesting additional complexity that had not been previously appreciated. This study has important clinical implications for the development of diagnostic assays to detect ETS rearranged prostate cancer. Incorporation of these less common ERG rearranged prostate cancer fusion assays could further increase the sensitivity of the current PCR-based approaches.
  Modern Pathology 04/2010; 23(4):539-46. · 4.79 Impact Factor
• Article: Outcome prediction for prostate cancer detection rate with artificial neural network (ANN) in daily routine.

  Thorsten H Ecke, Peter Bartel, Steffen Hallmann, Stefan Koch, Jürgen Ruttloff, Henning Cammann, Michael Lein, Mark Schrader, Kurt Miller, Carsten Stephan
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  ABSTRACT: We evaluated the use of the artificial neural network (ANN) program "ProstataClass" of the Department of Urology and the Institute of Medical Informatics at the Charité-Universitätsmedizin Berlin in daily routine to increase prostate cancer (CaP) detection rate and to reduce unnecessary biopsies. From May 2005 to April 2007, a total of 204 patients were included in the study. The Beckman Access PSA assay was used, and pretreatment prostate specific antigen (PSA) was measured prior to digital rectal examination (DRE) and 12 core systematic transrectal ultrasound (TRUS) guided biopsies. The individual ANN predictions were generated with the use of the ANN application for the Beckman Access PSA and free PSA assays, which relies on age, PSA, percent free prostate specific antigen (%fPSA), prostate volume, and DRE. Diagnostic validity of total prostate specific antigen (tPSA), %fPSA, and the ANN was evaluated by ROC curve analysis. PSA and %fPSA ranged from 4.01 to 9.91 ng/ml (median: 6.65) and 5% to 48% (median: 15%), respectively. Of all men, 46 (22.5%) demonstrated suspicious DRE findings. Total prostate volume ranged from 7.1 to 119.2 cc (median: 35). Overall, 71 (34.8%) CaP were detected. Of men with suspicious DRE, 28 (60.9%) had CaP on initial biopsy. The ANN was 78% accurate in the original report. The AUC of ROC curve analysis was 0.51 for PSA, 0.66 for %PSA, and 0.72 for the ANN-Output, respectively. Our results in this independent cohort show that ANN is a very helpful parameter in daily routine to increase the CaP detection rate and reduce unnecessary biopsies.
  Urologic Oncology 04/2010; 30(2):139-44. · 3.22 Impact Factor
• Article: Prevalence of TMPRSS2–ERG and SLC45A3–ERG gene fusions in a large prostatectomy cohort

  Raquel Esgueva, Sven Perner, Christopher J LaFargue, Veit Scheble, Carsten Stephan, Michael Lein, Florian R Fritzsche, Manfred Dietel, Glen Kristiansen, Mark A Rubin
  [show abstract] [hide abstract]
  ABSTRACT: The majority of prostate cancers harbor recurrent gene fusions between the hormone-regulated TMPRSS2 and members of the ETS family of transcription factors, most commonly ERG. Prostate cancer with ERG rearrangements represent a distinct sub-class of tumor based on studies reporting associations with histomorphologic features, characteristic somatic copy number alterations, and gene expression signatures. This study describes the frequency of ERG rearrangement prostate cancer and three 5 prime (5′) gene fusion partners (ie, TMPRSS2, SLC45A3, and NDRG1) in a large prostatectomy cohort. ERG gene rearrangements and mechanism of rearrangement, as well as rearrangements of TMPRSS2, SLC45A3, and NDRG1, were assessed using fluorescence in situ hybridization (FISH) on prostate cancer samples from 614 patients treated using radical prostatectomy. ERG rearrangement occurred in 53% of the 540 assessable cases. TMPRSS2 and SLC45A3 were the only 5′ partner in 78% and 6% of these ERG rearranged cases, respectively. Interestingly, 11% of the ERG rearranged cases showed concurrent TMPRSS2 and SLC45A3 rearrangements. TMPRSS2 or SLC45A3 rearrangements could not be identified for 5% of the ERG rearranged cases. From these remaining cases we identified one case with NDRG1 rearrangement. We did not observe any associations with pathologic parameters or clinical outcome. This is the first study to describe the frequency of SLC45A3–ERG fusions in a large clinical cohort. Most studies have assumed that all ERG rearranged prostate cancers harbor TMPRSS2–ERG fusions. This is also the first study to report concurrent TMPRSS2 and SLC45A3 rearrangements in the same tumor focus, suggesting additional complexity that had not been previously appreciated. This study has important clinical implications for the development of diagnostic assays to detect ETS rearranged prostate cancer. Incorporation of these less common ERG rearranged prostate cancer fusion assays could further increase the sensitivity of the current PCR-based approaches.Keywords: prostate cancer; ETS rearrangements; prevalence
  Modern Pathology 01/2010; 23(4):539-546. · 4.79 Impact Factor
• Article: Gene promoter methylation and its potential relevance in early prostate cancer diagnosis.

  Isabel Steiner, Klaus Jung, Philipp Schatz, Torsten Horns, Daniel Wittschieber, Michael Lein, Manfred Dietel, Andreas Erbersdobler
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  ABSTRACT: We investigated hypermethylation of the glutathione S-transferase pi (GSTP1), retinoic acid receptor β2 (RARβ2), adenomatous polyposis coli (APC) and paired-like homeodomain transcription factor 2 (PITX2) gene promoters which could serve as a sensitive tool to indicate a risk of prostate cancer even in histologically tumor-free tissues. Tumor tissues and non-neoplastic tissues at variable distances from the tumor foci were retrieved from 25 formalin-fixed and paraffin-embedded prostatectomy specimens and subjected to DNA extraction. The methylation levels were assessed by means of different assay technologies. Significantly increased methylation levels in cancer specimens were found for all promoter regions (GSTP1: 21/25, 84%; RARβ2: 24/25, 96%; APC: 21/25, 84%; PITX2: 20/25, 80%) and in most samples containing prostatic intraepithelial neoplasia. Several samples showed increased RARβ2 and APC methylation in adjacent non-neoplastic tissue. An association between the methylation extent of GSTP1, APC and RARβ2, respectively, and primary Gleason grade was detectable. GSTP1 methylation was also associated with extraprostatic tumor extension. GSTP1, APC, RARβ2 and PITX2 methylation occur frequently in prostate cancer, making these markers sensitive tools for the detection of neoplastic lesions in the prostate. For RARβ2, the results suggest a kind of methylation field effect which could be helpful for the detection of prostate cancer. Larger studies are necessary to investigate a potential correlation of GSTP1, RARβ2 and APC hypermethylation with tumor aggressiveness.
  Pathobiology 01/2010; 77(5):260-6. · 1.18 Impact Factor
• Article: Prostate specific antigen density to predict prostate cancer upgrading in a contemporary radical prostatectomy series: a single center experience.

  Ahmed Magheli, Stefan Hinz, Claudia Hege, Carsten Stephan, Klaus Jung, Kurt Miller, Michael Lein
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  ABSTRACT: We investigated the value of pretreatment prostate specific antigen density to predict Gleason score upgrading in light of significant changes in grading routine in the last 2 decades. Of 1,061 consecutive men who underwent radical prostatectomy between 1999 and 2004, 843 were eligible for study. Prostate specific antigen density was calculated and a cutoff for highest accuracy to predict Gleason upgrading was determined using ROC curve analysis. The predictive accuracy of prostate specific antigen and prostate specific antigen density to predict Gleason upgrading was evaluated using ROC curve analysis based on predicted probabilities from logistic regression models. Prostate specific antigen and prostate specific antigen density predicted Gleason upgrading on univariate analysis (as continuous variables OR 1.07 and 7.21, each p <0.001) and on multivariate analysis (as continuous variables with prostate specific antigen density adjusted for prostate specific antigen OR 1.07, p <0.001 and OR 4.89, p = 0.037, respectively). When prostate specific antigen density was added to the model including prostate specific antigen and other Gleason upgrading predictors, prostate specific antigen lost its predictive value (OR 1.02, p = 0.423), while prostate specific antigen density remained an independent predictor (OR 4.89, p = 0.037). Prostate specific antigen density was more accurate than prostate specific antigen to predict Gleason upgrading (AUC 0.61 vs 0.57, p = 0.030). Prostate specific antigen density is a significant independent predictor of Gleason upgrading even when accounting for prostate specific antigen. This could be especially important in patients with low risk prostate cancer who seek less invasive therapy such as active surveillance since potentially life threatening disease may be underestimated. Further studies are warranted to help evaluate the role of prostate specific antigen density in Gleason upgrading and its significance for biochemical outcome.
  The Journal of urology 11/2009; 183(1):126-31. · 4.02 Impact Factor
• Article: Internal validation of an artificial neural network for prostate biopsy outcome.

  Carsten Stephan, Henning Cammann, Martin Bender, Kurt Miller, Michael Lein, Klaus Jung, Hellmuth-A Meyer
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  ABSTRACT: To carry out an internal validation of the retrospectively trained artificial neural network (ANN) 'ProstataClass'. A prospectively collected database of 393 patients undergoing 8-12 core prostate biopsy was analyzed. Data of these patients were applied to the online available ANN 'ProstataClass' using the Elecsys total prostate-specific antigen (tPSA) and free PSA (fPSA) assays. Beside the internal validation of the ANN 'ProstataClass' an additional ANN (named as ANN internal validation: ANNiv) only using the 393 prospective patient data was evaluated. The new ANN model was constructed with the MATLAB Neural Network Toolbox. Diagnostic accuracy was evaluated by receiver operator characteristic (ROC) curves comparing the areas under the ROC curves (AUC) and specificities at 90% and 95% sensitivity. Within a tPSA range of 1.0-22.8 ng/mL, 229 men (58.3%) had prostate cancer (PCa). tPSA, %fPSA and the number of positive digital rectal examinations (DRE) differed significantly from the cohort of patients of the ANN 'ProstataClass', whereas age and prostate volume were comparable. AUCs for tPSA, %fPSA and the ANN 'ProstataClass' were 0.527, 0.726 and 0.747 (P = 0.085 between %fPSA and ANN). The AUC of the ANNiv (0.754) was significantly better compared with %fPSA (P = 0.021), whereas the AUC of two ANN models built on external cohorts (0.726 and 0.729) showed no differences to %fPSA and the other ANN models. Significant differences of DRE status and %fPSA medians decrease the power of the 'ProstataClass' ANN in the internal validation cohort. The effect of retrospective data evaluation the 'ProstataClass' cohort and prospective fPSA measurement may be responsible for %fPSA differences. All ANN models built with different PSA and fPSA assays performed equally if applied to the two cohorts.
  International Journal of Urology 11/2009; 17(1):62-8. · 1.75 Impact Factor